Waveguide detection of right-angle-scattered light in flow cytometry

• US 6,154,276 A
• Filed: 02/23/1998
• Issued: 11/28/2000
• Est. Priority Date: 02/23/1998
• Status: Expired due to Fees

• Current Assignee: Lawrence Livermore National Security LLC (Government of the United States of America)
• Original Assignee: Regents of the University of California (University of California)
• Inventors: Mariella, Raymond P. Jr.
• Primary Examiner(s): Font, Frank G.
• Assistant Examiner(s): STAFIRA, MICHAEL PATRICK
• Application Number:
  US09/027,764
• Time in Patent Office: 1,009 Days
• Field of Search: 356/336, 356/337, 356/335, 356/410, 356/246, 356/440
• US Class Current: 356/337
• CPC Class Codes: G01N 15/1436 the optical arrangement forming an integrated apparatus with the sample container, e.g. a flow cell

Abstract

A transparent flow cell is used as an index-guided optical waveguide. A detector for the flow cell but not the liquid stream detects the Right-Angle-Scattered (RAS) Light exiting from one end of the flow cell. The detector(s) could view the trapped RAS light from the flow cell either directly or through intermediate optical light guides. If the light exits one end of the flow cell, then the other end of the flow cell can be given a high-reflectivity coating to approximately double the amount of light collected. This system is more robust in its alignment than the traditional flow cytometry systems which use imaging optics, such as microscope objectives.

Claims

1. A flow cytometer flow cell, comprising:

a transparent flow cell having a hollow flow capillary to allow the flow of a sheath liquid therethrough, wherein said transparent flow cell comprises a solid-state material having an index of refraction that is greater than that of said sheath liquid, wherein particles suspended in said sheath liquid will produce scattered light from a laser beam directed at said particles, wherein a first portion of said scattered light will propagate into said solid-state material of said transparent flow cell; and

a detector positioned to detect a second portion of said first portion of said scattered light.

2. The flow cytometer flow cell of claim 1, wherein said detector further comprises a hole, wherein said detector is coupled to an end of said flow cell.

3. The flow cytometer flow cell of claim 1, further comprising a laser to provide a laser beam to be directed at said particles suspended in said sheath liquid.

4. The flow cytometer flow cell of claim 3, wherein said laser comprises a diode laser.

5. The flow cytometer flow cell of claim 2, wherein said transparent flow cell further comprises a high-reflection coating on the opposite end from said detector.

6. The flow cytometer flow cell of claim 2, wherein said transparent flow cell further comprises an anti-reflection coating on the same end as said detector.

7. The flow cytometer flow cell of claim 1, further comprising means for providing a liquid for input into said hollow flow capillary for production of said liquid sheath.

8. The flow cytometer flow cell of claim 1, wherein said transparent flow cell further comprises a cylindrical shape with a first end and a second end with a cylinder portion therebetween, wherein said cylinder portion comprises high-reflector material thereover except for a hole or absence of said high-reflector material thereon to allow passage of a laser beam into said transparent flow cell.

9. The flow cytometer flow cell of claim 8, wherein said first end comprises a high-reflection coating.

10. The flow cytometer flow cell of claim 8, wherein said second end comprises an anti-reflection coating, wherein said detector is coupled to said second end and said anti-reflection coating.

11. The flow cytometer flow cell of claim 2, wherein said hollow flow capillary exits said transparent flow cell at a location other than at the interface between said transparent flow cell and said detector.

12. The flow cytometer flow cell of claim 1, wherein a length of said hollow flow capillary is formed along the longitudinal axis of said transparent flow cell, wherein a second length of said hollow flow capillary is formed from an end of said first length toward the outer wall of and forming an opening in said flow cell, wherein said liquid sheath can move through said flow cell within said hollow flow capillary along said longitudinal axis and exit out of the side of said transparent flow cell.

13. The flow cytometer flow cell of claim 12, wherein said detector is coupled to an end of said flow cell.

14. The flow cytometer flow cell of claim 12, further comprising a laser to provide a laser beam to be directed at said particles suspended in said sheath liquid.

15. The flow cytometer flow cell of claim 14, wherein said laser comprises a diode laser.

16. The flow cytometer flow cell of claim 12, wherein said transparent flow cell further comprises a high-reflection coating on the opposite end from said detector.

17. The flow cytometer flow cell of claim 12, wherein said transparent flow cell further comprises an anti-reflection coating on the same end as said detector.

18. The flow cytometer flow cell of claim 11, further comprising means for providing a liquid for input into said hollow flow capillary for production of said liquid sheath.

19. The flow cytometer flow cell of claim 11, wherein said transparent flow cell further comprises a cylindrical shape with a first end and a second end with a wrapped plane portion therebetween, wherein said wrapped plane portion comprises high-reflector material thereover except for a hole or absence of said high-reflector material thereon to allow passage of a laser beam into said transparent flow cell.

20. The flow cytometer flow cell of claim 19, wherein said first end comprises a high-reflection coating.

21. The flow cytometer flow cell of claim 19, wherein said second end comprises an anti-reflection coating, wherein said detector is coupled to said second end and said anti-reflection coating.

22. The flow cytometer flow cell of claim 1, further comprising a high reflector surface embedded within said transparent flow cell, said high reflector surface having a hole to allow the flow of said sheath liquid through said hollow flow capillary, wherein said high reflector surface is configured to reflect said second portion of scattered light out of said transparent flow cell and onto said detector.

23. The flow cytometer flow cell of claim 22, wherein said detector is coupled to said flow cell where said second portion of said scattered light exits said transparent flow cell.

24. The flow cytometer flow cell of claim 22, further comprising a laser to provide a laser beam to be directed at said particles suspended in said sheath liquid.

25. The flow cytometer flow cell of claim 24, wherein said laser comprises a diode laser.

26. The flow cytometer flow cell of claim 22, wherein said transparent flow cell further comprises a high-reflection coating on a first end.

27. The flow cytometer flow cell of claim 22, wherein said transparent flow cell further comprises a high-reflection coating on a second end.

28. The flow cytometer flow cell of claim 22, further comprising means for providing a liquid for input into said hollow flow capillary for production of said liquid sheath.

29. The flow cytometer flow cell of claim 22, further comprising an anti-reflection coating on the surface of said transparent flow cell where light exits said transparent flow cell.

30. The flow cytometer flow cell of claim 22, wherein said transparent flow cell further comprises a cylindrical shape with a first end and a second end with a wrapped plane portion therebetween, wherein said wrapped plane portion comprises high-reflector material thereover except for a first hole or absence of said high-reflector material thereon to allow passage of a laser beam into said transparent flow cell and a second hole or absence of said high-reflector material thereon to allow passage of a laser beam out of said transparent flow cell.

31. The flow cytometer flow cell of claim 30, wherein said first end comprises a high-reflection coating.

32. The flow cytometer flow cell of claim 30, wherein said second end comprises a high-reflection coating.

33. The flow cytometer flow cell of claim 30, further comprising an anti-reflection coating on the surface of said transparent flow cell where light exits said transparent flow cell.

34. The flow cytometer flow cell of claim 1, further comprising means for sorting cells that exit said hollow flow capillary.

35. The flow cytometer flow cell of claim 12, further comprising means for sorting cells that exit said hollow flow capillary.

36. The flow cytometer flow cell of claim 22, further comprising means for sorting cells that exit said hollow flow capillary.

37. The flow cytometer flow cell of claim 1, further comprising an extended light source to provide light to be directed at said particles suspended in said sheath liquid.